Diphenic acid monoamides

ABSTRACT

The present invention provides various novel diphenic acid monoamide compounds, novel pharmaceutical compositions and methods of use thereof, as well as novel methods of synthesis therefor. The novel diphenic acid monoamides of the present invention are leukotriene antagonists, 5-lipoxygenase inhibitors, and mediator release inhibitors providing activity useful for treating asthma, allergies, cardiovascular diseases, migraines, and immunoinflammatory conditions.

BACKGROUND OF THE INVENTION

The present invention provides various novel diphenic acid monoamidecompounds, novel pharmaceutical compositions, and methods of usethereof, as well as, novel methods of synthesis therefor. The noveldiphenic acid monoamides of the present invention are leukotrieneantagonists, 5-lipoxygenase inhibitors, and mediator release inhibitorsproviding activity useful for treating asthma, allergies, cardiovasculardiseases, migraines, and immunoinflammatory conditions.

It is disclosed in U.S. Pat. No. 3,995,045 that2'-[(3,6-dihydro-4-phenyl-1(2H)-pyridinyl orphenyl-1-piperidinyl)alkyl-aminocarbonyl] [1,1'-diphenyl]-2-carboxylicacids are useful as antiinflammatory agents and, further, are useful inthe prevention and inhibition of granuloma formation. Among thedifferences between the invention in U.S. Pat. No. 3,995,045 and thepresent invention are notably the presence of a pyridinyl or piperidinylring attached to an alkyl of an alkylaminocarbonyl group as well asvarious combination of substituents of aryls all of which are not thepresent invention.

SUMMARY OF THE INVENTION

The present invention is a compound of Formula I wherein R₁ is hydrogenor lower alkyl of from one to four carbons, inclusive; B is (a)--(CH₂)_(m) --, (b) --C(O)--CH═CH--, (c) --C(O)--(CH₂)_(q) --, (d)--(CH₂)_(q) (O)--, (e) --O--(CH₂)_(q), (f) --O--(CH₂)_(m) --O--, (g)--CH(OH) (CH₂)_(q) --, (h) --(CH₂)_(q) CH(OH)--, (i) --CH═CH--C(O)--,(j) --(CH═CH)_(m) --, (k) --(CH₂)_(m) O-- wherein m is an integer fromone to six; q is an integer of zero to six; and Z is a substituenthaving the structure as shown in the formula page as (a) i, (b) ii, (c)iii, (d) iv, (e) v, (f) vi, (g) vii, or (h) viii, wherein n is aninteger of zero to five and R may be the same or different and loweralkyl of from one to four carbons, inclusive; hydroxy, alkoxy of fromone to four carbons, inclusive, halogen, or Z as defined above exceptthat R is hydrogen; with the over all proviso that R may only beattached to a carbon and that each carbon may carry only one R; or apharmaceutically acceptable salt thereof.

The present invention also includes a pharmaceutical compositioncomprising an amount of a compound of Formula I wherein R₁, B, m, Z, n,q, and R are as defined above effective for treating asthma, allergies,cardiovascular disorder, migraine, or an immunoinflammatory condition,in admixture with a pharmaceutically acceptable carrier.

Further, the invention is a method of treating asthma, allergies,cardiovascular disorders, migraine, or an immunoinflammatory conditionby administering an antiasthma, antiallergy, cardiovascular,antimigraine, or antiimmunoinflammatory effective amount to a subject,including a human suffering therefrom, of a compound of Formula Iwherein R₁, B, m, Z, n, q, and R are as defined above or apharmaceutically acceptable salt thereof.

Finally, the present invention is also a novel process for thepreparation of a compound of Formula I wherein R₁, B, m, Z, n, q, and Rare as defined above which comprises reacting a compound of Formula IIIwherein R₁, B, m, Z, n, q, and R are as defined above with a compound ofFormula II to obtain the compound I (see Scheme I).

DETAILED DESCRIPTION OF INVENTION

The term "lower alkyl of from one to four carbons, inclusive" as usedherein includes methyl, ethyl, propyl, or butyl, and isomers thereof.

The term "lower alkoxy of from one to four carbons, inclusive" as usedherein includes methoxy, ethoxy, propoxy, or butyloxy, and isomersthereof.

The term "halogen" as used herein includes, Br, Cl, F, or CF₃.

Preferred compounds of the present invention are compounds of theFormula I wherein B is attached in either the meta or para position andthen B is (a) --CH₂)_(m) --, (c) --C(O)--(CH₂)_(q) --, (e)--O--(CH₂)_(q) --, or (f) --O--(CH₂)_(m) --O--.

More preferred compounds of the present invention are the preferredcompounds of Formula I wherein B is (a) --(CH₂)_(m) -- wherein m is onethrough three and Z is the moiety shown as (i) wherein n is 0 or n istwo when R is halo or alkyl of from one to four carbons, inclusive;wherein B is (c) --C(O)--(CH₂)_(q) -- wherein q is one through three andZ is the moiety shown as (i) wherein n is zero or n is two when R ishalo or alkyl of from one to four carbons, inclusive; wherein B is (e)--O--(CH₂)_(q) -- wherein q is one through three and Z is the moietyshown as (i) wherein n is zero or n is two when R is halo or alkyl offrom one to four carbons-, inclusive; and wherein B is (f)--O--(CH₂)_(m) --O-- wherein m is two or three and Z is the moiety shownas (i) wherein n is zero or n is two when R is halo or alkyl of from oneto four carbons, inclusive.

The most preferred compounds of Formula I are the more preferredcompounds wherein B is (a) wherein m is three and n is zero or n is twowhen R is chloro or methyl; B is (c) wherein q is three and n is zero orn is two when R is chloro or methyl; B is (e) wherein q is three and nis zero or n is two when R is chloro or methyl; and B is (f) wherein mis two and n is zero or n is two when R is chloro or methyl.

The pharmacologically acceptable salts of the present invention may bethose readily prepared with inorganic and organic bases, such as NaOH,KOH, Mg(OH)₂, Ca(OH)₂, NH₄ OH, substituted ammonium salts, L-arginine,choline, N-methyl glucamine and the like.

By virtue of the activity of the compounds having the Formula I of thepresent invention as leukotriene D4 antagonists, and inhibitors of5-lipoxygenase and histamine release from basophils the compounds areuseful in treating asthmas and allergies as well as cardiovasculardisorders, migraine, and immunoinflammatory conditions. See B.Samulesson, "Leukotrienes: Mediators of Immediate HypersensitivityReactions and Inflammation, "Science" Vol. 220, p 568 (1983); P. J.Piper, "Leukotrienes," Trends in Pharmaceutic Sciences, pp 75 & 77(1983), and J. L. Romson, et al, "Reduction of the Extent of IschemicMyocardial Injury by Neutrophil Depletion in the Dog," Circulation, Vol.67, pp 1016 (1983).

Additionally, the activity, of the compounds having the Formula I of thepresent invention is determined by the well known leukotriene receptorbinding assay that is described by R. F. Bruns, W. J. Thomsen and T. A.Pugsley in Life Sciences, 33, 645 (1983) or the Herxheimer in vivoantiallergy test described in H. Herxheimer, J. Physiol. (London), Vol.177, p. 251 (1952).

The antiasthma and antiallergic activity provides methods of treatmentfor hypersensitivity reaction having broad symptoms. For example, thesymptons may include dermatitis, lacrimation, nasal discharge, coughing,sneezing, nausea, vomiting, diarrhea, difficulty in breathing, pain,inflammation, and in severe cases, anaphylactic shock and circulatorycollapse. The symptoms may be found in man as well as other animalssuffering from bronchial asthma, seasonal pollinosis (e.g., hayfever),allergic rhinitis, urticoria, allergic conjunctivitis, food allergies,and anaphylactoid reactions.

Likewise, the activity of the compounds of Formula I provides a methodof treatment for cardiovascular disorders, particularly ischemia andmyocardial infarctions. The symptoms of a subject having acardiovascular disorder may be determined by special diagnosticprocedures directed to subjects having a history, general physicalappearance, and then detailed deviations from normal appearancessuggesting a cardiovascular disorder. Such disorders are also found inman as well as other mammals. Symptoms of the disorders are describedextensively in The Merck Manual 14th ed, (1982).

Further, method of treatment is provided by the compounds of Formula Iherein for migraine and immunoinflammation. The symptoms requiringtreatment for these purposes are readily recognized, particularly formigraine in man and/or immunoinflammation in man as well as othermammals.

Pharmaceutical compositions which also are the present invention areprepared from the compound of Formula I and salts thereof described asthe present invention having inert pharmaceutical carriers. Thecompositions may be either solid or liquid.

A physician or veterinarian of ordinary skill readily determines asubject who is exhibiting symptoms described herein. Regardless of theroute of administration selected, the compounds of the present inventionare formulated into pharmaceutically acceptable dosage forms byconventional methods known to the pharmaceutical art.

The compounds can be administered in such oral unit dosage forms such astablets, capsules, pills, powders, or granules. They also may beadministered rectally or vaginally in such forms as suppositories orbougies; they may also be introduced parenterally (e.g., subcutaneously,intravenously, or intramuscularly), using forms known to thepharmaceutical art. They are also introduced directly to an affectedarea (e.g., in the form of eye drops or by inhalation). For thetreatment of asthma or allergies such as erythema, the compounds of thepresent invention may also be administered topically in the form ofointments, creams, gels, or the like.

An effective but nontoxic quantity of the compound is employed intreatment. The ordinarily skilled physician or veterinarian will readilydetermine and prescribe the effective amount of the compound to preventor arrest the progress of the condition for which treatment isadministered. In so proceeding, the physician or veterinarian couldemploy relatively low dosages at first, subsequently increasing the doseuntil a maximum response is obtained.

Initial dosages of the compounds of the invention having Formula I areordinarily in the area of 1.0 mg/kg up to 500.0 mg/kg per day orally,preferably 5.0 mg/kg to 10.0 mg/kg per dose orally, given from one tofour times daily or as needed. When other forms of administration areemployed equivalent doses are administered.

Generally, the compounds, having the Formula I wherein R₁, B, m, Z, n, qand R are as defined above are prepared by stirring a compound ofFormula III wherein R₁, B, m, Z, n, q and R are as defined above withdiphenic acid anhydride in an inert organic solvent, such asdichloromethane, tetrahydrofuran, toluene, 1,2-dichloroethane, and thelike at about room temperature or at reflux for from 30 minutes toovernight, preferably less than five hours (See Scheme I).

The synthesis of the various anilines of the Formula III as definedabove are carried out using the procedures analogous to those describedeither by E. P. Kohler and H. M. Chadwell, Organic Synthesis Coll. Vol.1, 78 (1941) and H. F. Hodson, J. W. T. Lowlands, and C. M. Leaver,EPO28305 (1979) or G. Markl and A. Merz, Synthesis, 295 (1973), andSynthetic Communications (1976), 6, 53, or a modification of thesemethods. Nitrations were done according to the procedures of G. Powelland F. R. Johnson, Org. Syn., Coll, Vol. II, p 44, or R. E. Buckles andM. P. Bellis, Org. Syn., Coll. Vol. IV, p 722.

Method A (Scheme II₁ and II₂)

Generally, the preparation of the compounds of Formula III₁ or III₂wherein R and n are as defined above is the method as shown in SCHEMESII₁ or II₂. The necessary benzyl triphenylphosphonium halide salts ofFormula IV₁ or IV₂ wherein R and n are as defined above are prepared bythe procedure analogous to that described by R. Ketchan, D. Jambotkar,and L. Martenelli, J. Org. Chem., 27, 466 (1962) and Synthetic Commun.,6, 53 (1976). The aldehydes are either purchased or prepared by acondensation of acetaldehyde and a substituted benzaldehyde giving thenecessary cinnamaldehyde of Formula V₁ wherein R and n are as definedabove and Formula V₂. The aromatic aldehyde V₁ or V₂ andbenzyltriphenylphosphonium salt IV₁ or IV₂ are coupled usingn-butyllithium as the base in THF or toluene at a temperature of from-10° C. to reflux, preferably at room temperature. Reduction of thenitro group and double bond in the coupled product is accomplishedsimultaneously using about 50 psig hydrogen and a Raney Nickel catalystunder standard hydrogenation conditions.

Method B

Generally, the necessary aryl- or aralkyl-phenylketones of Formula XII₄,wherein R and n are defined above used as starting materials areprepared by obvious routes or as described below or purchased directly.

To prepare meta-substituted aniline side-chains the aryl or aralkylphenyl ketone XII₄ is nitrated directly at -15° to 0° C. in aceticanhydride and a nitric acid solution up to room temperature then reducedto the anilines in a solvent such as alcohol or ethers at roomtemperature and used directly or subjected to Wolff-Kischner reductionof the ketone (see The Merck Index, 10th Ed, pp ONR-97) and then reducedto the aniline.

The parasubstituted aniline side chains are prepared by performing aWolff-Kischner reduction (see The Merck Index, 10th Ed, pp ONR-97) onthe aryl- or aralkyl-phenylketones XII followed by nitration, isolationof the pure p-isomer and reduction of the nitro group to an anilineagain by a catalytic hydrogenation as cited above.

See Scheme III for preparation of both the meta- and para-substitutedanilines as described above.

Alternatively, the preparation of the compound of Formula III wherein Bis an alkyl of three carbons may be accomplished via a Claisen-Schmidtcondensation (see The Merck Index, 10th Ed, pp ONR-19) of anitroacetophenone and an aromatic aldehyde followed by reduction of thedouble-bond as accomplished above and the nitro group. The resultantketo-aniline could be used directly or the ketone could be removed by aWolff-Kischner reduction cited above. This preparation is shown inScheme IV.

Method C (see Scheme V and Scheme VI)

Generally, to prepare compounds of Formula III wherein B is--O--(CH₂)_(m) --0-- wherein m is as defined above; p-acetamido-phenolof Formula XXIII is condensed with a 1-bromo-δ-chloroalkane of FormulaXXII in alcohol-ether at room temperature. This intermediate of FormulaXXI is condensed with a substituted phenol of Formula XXI₁ wherein R andn is as defined above using NaH in DMF, THF, DMSO, or DMA to give theacetylated compound of Formula XX₆. This may be cleaved to the anilineusing 6N HCl. (See Scheme V).

To prepare compounds of Formula III wherein B is O--(CH₂)_(q) -- whereinq is defined as above; 4-fluoro nitrobenzene is condensed with aω-phenyl alkanol wherein q and R are defined as above in an ethersolvent at room temperature or at reflux preferably at reflux to givethe nitro compound of Formula X₅. This may be reduced to the aminocompound III₅ by the hydrogenation methods described above (see SchemeVI).

The products of the reactions described herein are isolated byconventional means such as crystallization, extraction, distillation,chromatography, and the like.

Preparation of the compounds of Formula I wherein R₁ is lower alkyl offrom one to four carbons, inclusive, is by methods analogous to thoseknown in the art from the compounds of Formula I wherein R₁ is hydrogen.

The salts; both inorganic and organic, including esters, of compounds ofFormula I described above are prepared by reacting the appropriate baseor alcohol with a stoichometric equivalent of the carboxylic acid groupson the compound of Formula I.

The compounds of this invention may also exist in hydrated or solvatedforms.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The process of this invention is further elaborated by therepresentative examples as follows.

PREPARATION OF INTERMEDIATES (See Schemes II₁ and II₂) PREPARATION 1-SEESCHEME II₁

a. 4-(Nitrophenyl)-butanal

In a round bottomed flask equipped with magnetic stirring and a nitrogenatmosphere was placed dichloromethane (500 ml) and 48.2 (0.128 mol) ofpyridinium dichromate. This slurry was stirred vigorously and 25.0 g of4-nitrophenylbutanol was added dropwise from a pressure equalizingaddition funnel. The reaction was stirred overnight. The resultant brownslurry was filtered through silica gel with diethylether and thesolution was concentrated to an oil by rotary-evaporation at aspiratorpressure. This oil was purified by column chromatography usingdichloromethane as eluent. A total of 15.4 g of the4-(nitrophenyl)butanal was obtained as a yellow oil.

b. 4-(5-Phenylpentyl)-aniline

In a round bottomed flask equipped with magnetic stirring, a nitrogenatmosphere, thermometer, and pressure equalizing addition funnel wasplaced 22.13 g of benzyltriphenylphosphonium chloride and 400 ml of drytetrahydrofuran. This slurry was cooled to <0° C. and 25.0 ml ofn-butyllithium (2.4M in hexane) was added dropwise while maintaining thetemperature at <5° C. After the addition of the n-butyllithium wascomplete, the slurry was stirred for six hours at room temperature then11.0 g (0.057 mol) of 4-nitrophenylbutanal was added dropwise and theresultant mixture was stirred overnight. The solvent was removed byrotary-evaporation and the residue was dissolved in hot toluene (1000ml) and filtered through silica gel and the silica gel was washed with3:1 cyclohexane:ethylacetate. The toluene and cyclohexane/ethyl acetatewas evaporated and the residue (10.2 g) was dissolved in 1/1 (v/v)tetrahydrofuran-methanol and 1.5 g of Raney-nickel was added. Thismixture was hydrogenated in a Parr™ shaker at three atmospheres ofhydrogen. After ˜15 hours a second 1.5 g portion of catalyst was addedand the hydrogenation was continued. When 95% of the theoretical amountof hydrogen had been taken up the reaction was stopped and the catalystwas removed by filtration through celite. After removal of the solventthe crude product was purified by column chromatography usingdichloromethane/cyclohexane. The 4-(5-phenylpentyl)aniline was obtainedas a tan oil after combination of the appropriate fractions and removalof the solvents. The yield was 5.77 g.

PREPARATION 2-SEE SCHEME II₁ or II₂

Benzeneamines may be prepared from known nitrobenzenes in a mannersimilar to the above preparation as follows.

4-[2-(3,4-Dichlorophenyl)ethyl]benzenamine

4-[2-(3,4-dichlorophenyl)ethenyl]nitrobenzene (34.0 g, 116 mmol) and 2 gof Raney nickel in 510 ml of tetrahydrofuran were shaken under hydrogenat 15 psi (103.4 kPascal) until four equivalents of hydrogen wereconsumed. The catalyst was removed by filtration, and the resultingfiltrate evaporated in vacuo to yield a residual tan solid.Recrystallization from hexane yielded 26.6 g (86%) of4-[2-(3,4-dichlorophenyl)ethyl]benzenamine, mp 72°-74° C.

PREPARATION 3-SEE SCHEME II₂

3,4-Dichlorocinnamaldehyde

This is prepared in a manner analogous to the method of U.S. Pat. No.4,375,475 as follows.

In a round bottomed flask equipped with magnetic stirring and a pressureequalizing addition funnel was placed 3,4-dichlorobenzaldehyde (140.0 g,0.8 mol) and acetaldehyde (300.0 ml, 5.36 mol) and the mixture wascooled to 5° C. Potassium hydroxide (5.1 g, 0.09 mol) was dissolved inmethanol (40.0 ml) and added dropwise to the solution of3,4-dichlorobenzaldehyde and acetaldehyde while maintaining thetemperature at ≦30° C. The resulting brown solution was stirred in anice bath for 30 minutes then acetic anhydride (400.0 ml) was added andthe reaction was heated to 100° C. for 30 minutes. After cooling thereaction to 30° C. a solution consisting of 102 ml concentrated HCl and1200 ml of water was added and the resulting mixture was heated toreflux for 30 minutes. The reaction as cooled to ice bath temperatureand the brown solid was collected and allowed to crystallize from hotcyclohexane after treatment with decolorizing charcoal. This gave 111.0g of 3,4-dichlorocinnamaldehyde, mp 100°-102° C.

The 3,4-dichlorocinnamaldehyde is then reacted with anitrobenzyltriphenylphosphonium chloride in a manner similar toPreparation 1 b above (see Scheme II₁). The resulting4-[4-(3,4-dichlorophenyl)butyl]benzamine is a compound of Formula IIIwherein R₁ is H, Z is 3,4-dichlorophenyl and B is butyl. The benzamineis for use as an intermediate as shown in Scheme I.

Other benzamines are prepared in a manner analogous to the followingpreparations which exemplify the Scheme VI and Scheme V.

PREPARATION I (SCHEME VI)

1-[4-(Aminophenoxy)]-2-phenyl ethane (III₅)

In a round bottom flask equipped with N₂ atmosphere, magnetic stirring,reflux condenser, heating mantle, and addition funnel was placed dry THF(50 ml) and sodium hydride (60% by water in oil, 1.6 g). To this slurrywas added phenethanol (4.32 g) dropwise followed by 4-fluoronitrobenzene(in 10 ml THF). The exothermic reaction was controlled with an ice bath.The red solution was stirred overnight at room temperature then refluxedfor four hours. The reaction was cooled and concentrated to dryness andthe residue was partitioned between CH₂ Cl₂ and H₂ O. The organic layerwas washed with H₂ O, dried, and evaporated to a light yellow oil. Thisoil was triturated with hexane to give a light orange solid (3.2 g)[Analysis C₁₄ H₁₃ NO₃, actual in parenthesis, (243.26); 69.12 (68.90);5.39 (5.39); 5.76 (5.71)]. The 1-(4-nitrophenoxy)-2-phenoxy)ethane ofFormula X₅ was reduced under the standard conditions to the aminoderivative. This low melting solid was analytically pure from thereaction after removal of the solvent [yield 2.5 g].

Analysis: C₁₄ H₁₅ NO (213.28); 78.84 (77.45); 7.09 (6.69); 6.57 (6.11)(actual in parenthesis).

PREPARATION II (SCHEME V)

1-[4-acetamidophenoxy]-3-chloropropane (XXI)

In a round bottomed flask equipped with magnetic stirring, refluxcondenser, and N₂ atmosphere was placed ethanol (500 ml), sodiumhydroxide (15.0 g), 4-acetamidophenol of Formula XXIII (30.0 g), and1-bromo-3-chloropropane of Formula XXII (47.0 g) and the mixture wasrefluxed overnight. Upon cooling off-white crystals were deposited andthen collected. The mother liquor was evaporated to dryness anddissolved in CH₂ Cl₂ then washed with H₂ O, dried, and concentrated to50 ml. A second crop of crystals were deposited. These were combinedwith the first crop of crystals and recrystallized from ethanol to givecolorless crystals (20.3 g), mp 128°-130° C.

PREPARATION III

1-(4-Acetamidophenoxy)-3-(3,4-dichlorophenoxy)propane XX₆

In a round bottomed flask equipped with magnetic stirring, refluxcondenser, N₂ atmosphere, and a heating mantle was placed DMF (80 ml)and to this was added cautiously NaH (60% by weight in oil, 1.76 g)followed by 3,4-dichlorophenol (7.15 g). This mixture was stirred forten minutes then 1-[4-acetamidophenoxy]-3-chloropropane of Formula XXI(10.0 g) was added; and the reaction mixture heated at 80° C. overnight.The mixture was cooled and the DMF removed in vacuo. The residue waspoured into ice water and the precipitated brown solid was collected byfiltration. The crude product was recrystallized from ethanol to givenan off white solid (9.8 g), mp 135°-7° C.

PREPARATION IV

1-(4-aminophenoxy)-3-(3,4-dichlorophenoxy)propane III₆

In a round bottomed flask equipped with magnetic stirring and a refluxcondenser was placed 50 ml of 6N HCL solution and1-(4-acetamidophenoxy)-3-(3,4-dichlorophenoxy)propane of Formula XX₆(9.5 g) and the slurry was refluxed overnight and then cooled. Theprecipitated tan solid was collected and triturated with ethyl acetateto give a solid (8.0 g).

Analysis: C₁₅ H₁₅ Cl₂ NO₂.HCl (348.66); 51.67, (51.93); 4.63 (4.76);4.02 (4.19); 30.51 (30.29) (actual in parenthesis).

The following anilines of Formula III are prepared in a manner analogousto the indicated method using appropriate starting materials.

    ______________________________________                                        Name                    mp      Method                                        ______________________________________                                        3-(2-(3,4-dichlorophenyl)ethyl)-aniline                                                               57-9    A                                             4-(3-(3,4-dichlorophenyl)propyl)-aniline                                                              oil     B                                             C.sub.15 H.sub.15 NCl.sub.2 (280.12)                                          Calcd:                                                                              C, 64.30; H, 5.39; N, 4.99; Cl 25.31                                    Found:                                                                              C, 63.77; H, 5.42; N, 5.07; Cl, 25.30                                   4-(3,4-dichlorobenzyl)-aniline                                                                        oil     B                                             C.sub.13 H.sub.11 NCl.sub.2 (252.14)                                          Calcd:                                                                              C, 61.93; H, 4.40; N, 5.56; Cl 28.12                                    Found:                                                                              C, 61.68; H, 4.55; N, 5.52                                              2-(2-(3,4-dichlorophenyl)ethyl-aniline                                                                101-3   A                                             4-(4-(3,4-dichlorophenyl)butyl)-aniline                                                               50-2    A                                             4-(3-phenylpropyl)aniline                                                     3-(3,4-dichlorobenzyl)-aniline                                                                        oil     B                                             C.sub.13 H.sub.11 NCl.sub.2 (252.14)                                          Calcd:                                                                              C, 61.93; H, 4.40; N, 5.56;                                             Found:                                                                              C, 62.17; H, 4.53; N, 5.59;                                             4-(4-phenylbutyl)-aniline                                                                             oil     A                                             C.sub.16 H.sub.19 N (225.33)                                                  Calcd:                                                                              C, 85.29; H, 8.50; N, 6.22                                              Found:                                                                              C, 85.40; H, 8.55; N, 6.20                                              4-(2-(3,4-dichlorophenyl)ethyl)-aniline                                                               72-4    A                                             4-(5-phenylpentyl)aniline                                                                             oil     A                                             ______________________________________                                    

Miscellaneous intermediates are prepared in a manner analogous to thedescription of the indicated method using appropriate startingmaterials.

    ______________________________________                                        Name                 mp °C.                                                                          Method                                          ______________________________________                                        1-phenyl-3-(3,4-dichlorophenyl)-                                                                   120-122  B                                               prop-2-enol                                                                   (1-benzoyl)-2-(3,4-dichlorophenyl)-                                           ethylene                                                                      3-(3,4-dichlorphenyl)propiophenone                                                                 44-45    B                                               3-nitro-3',4'-dichlorobenzophenone                                                                 133-5    B                                               1-phenyl-3-(3,4-dichlorophenyl)-                                                                   oil      B                                               propane                                                                       C.sub.15 H.sub.14 Cl.sub.2 (265.18)                                           Calcd:                                                                              C, 67.94; H, 5.32; Cl, 26.0                                             Found:                                                                              C, 68.29; H, 5.58; Cl 27.0                                              3,4-dichlorodiphenylmethane                                                                        oil      B                                               4-nitro-3',4'-dichlorodiphenyl-                                                                    95-7     B                                               methane                                                                       1-(4-nitrophenyl)-3-(3,4-dichloro-                                                                 72-4     B                                               phenyl)propane                                                                3-nitro-3',4'-dichlorostilbene                                                                     150-2    A                                               2-nitro-3',4'-dichlorostilbene                                                                     mixture  A                                               C.sub.14 H.sub.19 NO.sub.2 Cl.sub.2 (294.14)                                                       of iso-                                                  Calcd:                                                                              C, 57.17; H, 3.08; N, 4.76;                                                                      mers                                                       Cl, 24.0                                                                Found:                                                                              C, 56.95; H, 2.99; N, 4.77;                                                   Cl, 24.0                                                                1-(4-(nitrophenyl)-4-(3,4-dichloro-                                                                125-135  A                                               phenyl-1,3-butadiene                                                          C.sub.16 H.sub.11 NO.sub.2 Cl.sub.2 (320.175)                                 Calcd;                                                                              C, 60.02; H, 3.46; N, 4.37;                                                   Cl, 22.15                                                               Found:                                                                              C, 59.77; H, 3.47; N, 4.40;                                                   Cl, 22.39                                                               1-(4-nitrophenyl)-3-phenylprop-1-                                                                  oil      A                                               ene                                                                           C.sub.15 H.sub.13 NO.sub.2                                                    Calcd:                                                                              C, 75.30; H, 5.48; N, 5.85                                              Found:                                                                              C, 75.10; H, 5.70, N, 5.88                                               1-(4-nitrophenyl)-4-phenyl-1,3-                                                                   75-6     A                                               butadiene                                                                     ______________________________________                                    

EXAMPLE 1

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3,4-dichlorophenyl)propyl]phenyl]amino]carbonyl]-

In a round bottom flask equipped with magnetic stirring refluxcondenser, and heating mantle was placed diphenic acid anhydride (9.6 g,0.043 mol), 4-[3,4-dichlorophenylpropyl]benzamine (III₄) (12.0 g, 0.043mol), and 1,2-dichloroethane. The solution was refluxed for five hoursthen cooled to room temperature. The solution was evaporated to afoam-like material. The crude product was dissolved in hot 95% ethanolthen diluted with H₂ O (70 ml). After cooling an off-white solid wasdeposited. The product was collected and dried overnight at 60° C. athouse vacuum to yield 19.9 g of the title compound.

Analysis: C₂₉ H₂₃ Cl₂ NO₃ (504.41); 69.05 (69.33); 4.60 (4.78); 2.78(2.88); 14.06 (14.08).

EXAMPLE 2

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3-phenylpropoxy]phenyl]amino]carboxyl]-

In a round bottomed flask equipped with magnetic stirring, refluxcondenser, and heating mantle was placed 1,2-dichloroethane (30 ml),diphenic acid anhydride (0.74 g, 3.3 mmol), and4-(3-phenylpropyloxy)benzamine (0.75 g, 3.3 mmol) which had beenprepared from the corresponding hydrochloride salt in the followingmanner. The hydrochloride salt (1.0 g, 3.8 mmol) was slurried in waterand triethylamine (0.6 ml) was added followed by diethylether (20 ml).The ether layer was separated, dried, filtered, and evaporated to yield0.75 g of the desired benzamine. The reaction mixture was refluxed forthree hours then cooled to room temperature and concentrated to afoam-like material. The crude product was purified by columnchromatography on silica gel using dichloromethane and 9/1 v/vdichloromethane/methanol as eluents. The fractions containing theproduct were combined and evaporated to a foam-like material. The yieldwas 1.2 g (mp 98°-100° C.).

Analysis: C₂₉ H₂₅ NO₄ --0.05CH₃ OH (453.12 (451.52)); 77.14 (77.00);5.58 (5.61); 3.10 (3.09).

The following compounds of Formula I were prepared by a method analogousto either Example 1 or 2 using appropriate intermediates of Formula IIIwherein R₁, B, m, and R and n are defined in a corresponding manner.Each analysis shows the calculated values.

EXAMPLE 3

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(3,4-dichlorophenyl)ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₈ H₂₁ Cl₂ NO₃ (490.37); 68.58 (68.40); 4.42 (4.39); 2.85(2.95); 14.46 (14.44).

EXAMPLE 4

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[2-(3,4-dichlorophenyl)ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₈ H₂₁ NO₃ (490.37); 68.54 (68.37); 4.42 (4.46); 2.85 (2.77),14.46 (14.54).

EXAMPLE 5

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(3,4-dichlorophenyl)ethyl]phenyl]methylamino]carbonyl]-

Analysis: C₂₉ H₂₃ Cl₂ NO₃ --0.45 C₂ H₅ OH 504.44 (525.18); 68.38(68.37); 4.93 (4.84); 2.67 (2.70); 13.50 (13.52)

EXAMPLE 6

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[3-(3,4-dichlorophenyl)propyl]phenyl]amino]carbonyl]

Analysis: C₂₉ H₂₃ Cl₂ NO₃.0.25 H₂ O 508.91 (504.41); 68.44 (68.33); 4.65(4.58); 2.75 (2.75); 14.05 (13.93).

EXAMPLE 7

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(3,4-dimethoxyphenyl)ethyl]phenyl]amino]carbonyl]-

Analysis: C₃₀ H₂₇ NO₅ --0.38 C₆ H₁₂ 481.53 (513.51); 75.49 (75.21); 6.18(6.17); 2.73 (2.73).

EXAMPLE 8

[1,1-Biphenyl]-2-carboxylic acid,2'-[[[4-(4-chlorophenyl)ethyl]phenyl]methylamino]carbonyl]-

Analysis: C₂₉ H₂₄ ClNO₃ (469.97); 74.12 (74.30); 5.15 (5.20); 2.98(2.99); 7.54 (7.63).

mp 185°-7° C.

EXAMPLE 9

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[(3,4-dichlorophenyl)methyl]phenyl]amino]carbonyl]-

Analysis: C₂₇ H₁₉ Cl₂ NO₃ (476.36); 55.35 (55.15); 3.22 (3.47); 4.96(4.91); 25.13 (24.95).

mp 202°-4° C.

EXAMPLE 10

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[2-[4-(trifluoromethyl)phenyl]ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₉ H₂₂ F₃ NO₂ (489.51); 71.16 (70.82); 4.53 (4.79); 2.86(2.79); 11.64 (11.33).

mp 148°-50° C. EXAMPLE 11

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[2-[(4-chlorophenyl)ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₈ H₂₂ ClNO₃ (489.51); 73.76 (74.09); 4.86 (5.17); 3.07(2.95); 7.78 (7.85).

EXAMPLE 12

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[2-[2-(3,4-dichlorophenyl)ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₈ H₂₁ Cl₂ NO₃ (490.39); 57.17 (56.95); 3.08 (2.99); 4.76(4.77); 24.11 (24.61).

mp 193°-5° C.

EXAMPLE 13

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-[3,5-bis(trifluoromethyl)phenyl]ethyl]phenyl]amino]carbonyl]-

Mass Spectrum (M/Z) P+ 557, 539, 333, 180, 106.

IR KBr pellet cm⁻¹ 1654, 1606, 1548, 1382, 1280, 1176, 1134.

NMR (DMSO, δ), 12.78 (1H), 7.93-6.79 (15H), 3.00 (2H), 2.78 (2H).

EXAMPLE 14

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenyl]ethyl]phenyl]amino]carbonyl]-

Mass Spectrum M/Z P+ 549, 331, 313, 270, 225, 1197, 181, 152, 106.

IR KBr pellet cm⁻¹ 3400(b), 2958, 1702, 1603, 1539, 1437, 1235.

NMR (DMSO, δ), 8.25 (1H), 7.8-6.9 (14H), 5.03 (1H), 2.75 (4H), 1.39(18H).

EXAMPLE 15

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[4-(3,4-dichlorophenyl)butyl]phenyl]amino]carbonyl]-

Analysis: C₃₀ H₂₅ Cl₂ NO₃ (518.45); 69.50 (69.35); 4.86 (4.86); 2.70(2.69); 13.68 (13.92).

mp 170°-4° C.

EXAMPLE 16

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(3-phenylpropyl]phenyl]amino]carbonyl]-

Analysis: C₂₉ H₂₅ NO₃ (435.52); 79.98 (80.13); 5.79 (5.81); 3.22 (3.29).

EXAMPLE 17

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[(3,4-dichlorophenyl)methyl]phenyl]amino]carbonyl]-

Analysis: C₂₇ H₁₉ Cl₂ NO₃ --1.5 CH₃ OH 476.36 (524.42); 65.27 (65.36);4.80 (4.05); 2.67 (2.67); 13.52 (13.85).

EXAMPLE 18

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(4-methylphenyl)ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₉ H₂₅ NO₃ --0.8 C₂ H₅ OH 435.53 (472.38); 77.80 (77.96),6.78 (6.42); 2.96 (2.94).

mp 110°-2° C.

EXAMPLE 19

[1,1'-Biphenyl]-2-carboxylic acid, 2'-[[[4-(4-phenyl-butyl)phenyl]amino]carbonyl]-

Analysis: C₃₀ H₂₇ NO₃ (449.27); 80.15 (79.92); 6.05 (6.30); 3.12 (3.18).

mp 70°-74° C.

EXAMPLE 20

[1,1'-Biphenyl]-2-carboxylic acid, 2'-[[4-decylphenyl)-amino]carbonyl]-

Analysis: C₃₀ H₃₅ NO₃ --0.25 H₂ O 457.64 (462.12); 77.97 (77.97); 7.74(7.53); 3.03 (3.07).

mp 55°-60° C.

EXAMPLE 21

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(5-phenylpentyl)phenyl]amino]carbonyl]-

Analysis: C₃₁ H₂₉ NO₃ (463.55); 80.32 (80.18); 6.30 (6.32); 3.02 (2.96).

EXAMPLE 22

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(pentafluorophenyl)ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₈ H₁₈ F₅ NO₃ (511.45); 64.61 (64.58); 3.68 (3.74); 2.69(2.73); F, 18.25 (17.89).

EXAMPLE 23

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[2-[4-(1-oxo-3-phenyl-2-propenyl)phenyl]ethyl]amino]carbonyl]-

Analysis: C₃₁ H₂₅ NO₄ --0.2 H₂ O 475.52 (479.12); 77.70 (77.71); 5.34(5.34); 2.92 (2.82).

EXAMPLE 24

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[4-(2-furanyl)butyl]phenyl]amino]carbonyl]-

Analysis: C₂₈ H₂₅ NO₄ --0.17 H₂ O 439.49 (442.55); 75.99 (75.99); 5.77(5.94); 3.16 (3.10).

EXAMPLE 25

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(1-oxo-3-phenyl-2-propenyl)phenyl]amino]carbonyl]-

Analysis: C₂₉ H₂₁ NO₄ (447.50); 77.84 (77.54); 4.73 (4.90); 3.13 (3.06).

EXAMPLE 26

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[4-(3-pyridinyl)butyl]phenyl]amino]carbonyl]-

Analysis: C₂₉ H₂₆ N₂ O₃ (450.54); 72.78 (72.68); 5.57 (5.32); 5.78(5.85).

EXAMPLE 27

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3,4-dichlorophenyl)-1-oxopropyl]phenyl]amino]carbonyl-

Analysis: C₂₉ H₂₁ Cl₂ NO₄ --0.25 H₂ O 522.90 (518.40); 66.61 (66.34);4.14 (4.24); 2.68 (2.70).

EXAMPLE 28

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[3-(3,4-dichlorphenyl)-1-oxopropyl)phenyl]amino]carbonyl]-

Analysis: C₂₉ H₂₁ Cl₂ NO₄.0.13 C₂ H₄ Cl₂ 531.26 (518.40); 66.15 (66.09);4.08 (4.31); 2.64 (2.72); 15.08 (14.50).

EXAMPLE 29

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-[3,4-dichlorophenoxy]propoxy]phenyl]amino]carbonyl]-

Analysis: C₂₉ H₂₃ Cl₂ NO₅.0.15 CH₂ Cl₂ 549.15 (536.41); 63.75 (63.56);4.28 (4.44); 2.55 (2.55); 14.85 (14.75).

EXAMPLE 30

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(2,5-dimethylphenoxy]ethoxy]phenyl]amino]carbonyl]-

Analysis: C₃₀ H₂₇ NO₅.0.2 H₂ O 481.53 (485.13); 74.27 (74.38); 5.69(5.43); 2.89 (2.93).

EXAMPLE 31

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(3,4-dichlorophenoxy)ethoxy]phenyl]amino]carbonyl]-

Analysis: C₂₈ H₂₁ Cl₂ NO₅.0.5H₂ O 522.38 (531.39); 63.28 (63.23); 4.17(4.17); 2.64 (2.66); 13.34 (13.80).

EXAMPLE 32

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(2-phenoxyethoxy)phenyl]amino]carbonyl]

Analysis: C₂₈ H₂₃ NO₅.0.25 H₂ O 453.49 (458.00); 73.47 (73.43); 5.12(5.39); 3.06 (3.17).

EXAMPLE 33

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(3-phenoxypropoxy)phenyl]amino]carbonyl]

Analysis: C₂₉ H₂₅ NO₅.0.37 H₂ O 467.52 (473.81); 73.51 (73.45); 5.40(5.32); 2.96 (2.89).

EXAMPLE 34

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-(1-oxo-3-phenyl]propyl)phenyl]amino]carbonyl]-

Analysis: C₂₉ H₂₃ NO₄.0.78 H₂ O 449.51 (463.56); 75.15 (75.15); 5.34(5.06); 3.02 (2.97).

EXAMPLE 35

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[2-[2-[N-methylpyrrolyl]]ethenyl]phenyl]amino]carbonyl]-

Analysis: C₂₇ H₂₂ N₂ O₃.0.4 H₂ O 422.49 (429.70); 75.47 (75.44); 5.35(5.44); 6.52 (6.50).

EXAMPLE 36

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[2-[2-furyl]ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₆ H₂₁ NO₄ (411.40); 75.89 (75.58); 5.14 (5.37); 3.40 (3.38).

EXAMPLE 37

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[3-[(3,4-dimethoxyphenyl)-3-oxopropyl]phenyl]amino]carbonyl]-

Analysis: C₃₁ H₂₇ NO₆.0.5 C₆ H₁₂ 509.58 (551.66); 74.03 (74.11); 6.03(7.32); 2.32 (2.54).

EXAMPLE 38

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[3-(1,3-benzodioxol-5-yl)-1-oxopropyl]phenyl]amino]carbonyl]-

Analysis: C₃₀ H₂₃ NO₆ 493.49; 73.01 (72.95); 4.70 (4.90); 2.84 (2.86).

EXAMPLE 39

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[2-[(3,4-dihydroxyphenyl)ethyl]phenyl]amino]carbonyl]-

Analysis: C₂₈ H₂₃ NO₅.0.9 H₂ O 453.47 (469.74); 71.60 (71.66); 5.32(5.50); 2.98 (3.06).

EXAMPLE 40

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[3-[3,4-dimethoxyphenyl)-1-hydroxypropyl]phenyl]amino]carbonyl]-

Analysis: C₃₁ H₂₉ NO₆.0.5 H₂ O.0.5 C₂ H₆ O 511.59 (543.63); 70.70(70.98); 6.4 (5.87); 2.57 (2.56).

EXAMPLE 41

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[3-[(2,5-dimethoxyphenyl)-3-hydroxypropyl]phenyl]amino]carbonyl]-

Analysis: C₃₁ H₂₉ NO₆ 511.55; 72.78 (73.15); 5.71 (6.02); 2.74 (2.53).

EXAMPLE 42

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[1-[(phenyl-1-oxo-methyl]phenyl]amino]carbonyl]-

Analysis: C₂₇ H₁₉ NO₄.1.0 C₆ H₁₂ 421.43 (505.59); 78.39 (78.43); 6.18(7.03); 2.77 (2.45).

EXAMPLE 43

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[4-[(phenoxy)phenyl]amino]carbonyl]-

Analysis: C₂₆ H₇₉ NO₄.0.5 C₂ H₆ O 409.42 (432.46); 74.98 (74.95); 5.12(5.25); 3.24 (3.10).

Additionally, componds of Formula I are prepared by similar analogousmethods as described above in Examples 1 or 2 using appropriateintermediates of Formula III wherein R, B, m, R and n are defined in acorresponding manner.

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3,4-dimethoxyphenyl)propyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[4-(3,4-dimethoxyphenyl)]butyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[5-(3,4-dimethoxyphenyl)pentyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-(3,4-dimethoxyphenyl)methyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[2-(3,4-dimethoxyphenyl)ethyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[3-(3,4-dimethoxyphenyl)propyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[4-(3,4-dimethoxyphenyl)butyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[5-(3,4-dimethoxyphenyl)pentyl]phenyl]amino]carbonyl-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(3,4-dihydroxyphenyl)methyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carbboxylic acid,2'-[[[4-[2-(3,4-dihydroxyphenyl)ethyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3,4-dihydroxyphenyl)propyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[4-(3,4-dihydroxyphenyl)butyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[5-(3,4-dihydroxyphenyl)pentyl]phenyl]amino]carbonyl]-

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-(3,4-dihydroxyphenyl)methyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[2-(3,4-dihydroxyphenyl)ethyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[3-(3,4-dihydroxyphenyl)propyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[4-(3,4-dihydroxypheny)butyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2'-carboxylic acid,2'-[[[3-[5-(3,4-dihydroxyphenyl)pentyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-carboxylic acid,2'-[[[4-(3-hydroxy-4-methoxyphenyl)methyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(4-hydroxy-3-methoxyphenyl)methyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(3-hydroxy-4-methoxyphenyl)ethyl]phenyl]amino]-carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(4-hydroxy-3-methoxyphenyl)ethyl]phenyl]amino]carbonyl]-,

[1,1-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3-hydroxy-4-methoxyphenyl)propyl]phenyl]amino]carbonyl]-,

[1,1-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(4-hydroxy-3-methoxyphenyl)propyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[4-(3-hydroxy-4-methoxyphenyl)butyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[4-(4-hydroxy-3-methoxyphenyl)butyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[5-(3-hydroxy-4-methoxyphenyl)pentyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[5-(4-hydroxy-3-methoxyphenyl)pentyl]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3,4-dimethoxyphenyl)propoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[[4-(3,4-dimethoxyphenyl)]butyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[[5-(3,4-dimethoxyphenyl)pentyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-(3,4-dimethoxyphenyl)methoxyphenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[2-(3,4-dimethoxyphenyl)ethoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[3-(3,4-dimethoxyphenyl)propoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[[4-(3,4-dimethoxyphenyl)butyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[[5-(3,4-dimethoxyphenyl)pentyl]oxy]phenyl]amino]carbonyl-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(3,4-dihydroxyphenyl)methoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(3,4-dihydroxyphenyl)ethoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3,4-dihydroxyphenyl)propoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[[4-(3,4-dihydroxyphenyl)butyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[[5-(3,4-dihydroxyphenyl)pentyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-(3,4-dihydroxyphenyl)methoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[2-(3,4-dihydroxyphenyl)ethoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[3-(3,4-dihydroxyphenyl)propoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[3-[[4-(3,4-dihydroxyphenyl)butyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2'-carboxylic acid,2'-[[[3-[[5-(3,4-dihydroxyphenyl)pentyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(3-hydroxy-4-methoxyphenyl)methoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-(4-hydroxy-3-methoxyphenyl)methoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(3-hydroxy-4-methoxyphenyl)ethoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[2-(4-hydroxy-3-methoxyphenyl)ethoxy]phenyl]amino]carbonyl]-,

[1,1-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3-hydroxy-4-methoxyphenyl)propoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(4-hydroxy-3-methoxyphenyl)propoxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[[4-(3-hydroxy-4-methoxyphenyl)butyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[[4-(4-hydroxy-3-methoxyphenyl)butyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[[5-(3-hydroxy-4-methoxyphenyl)pentyl]oxy]phenyl]amino]carbonyl]-,

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[[5-(4-hydroxy-3-methoxyphenyl)pentyl]oxy]phenyl]amino]carbonyl]-.

Likewise, using the above generally described procedures the followingcompounds as prepared.

1,1'-Biphenyl-2-carboxylic acid,2'-[[[4-[3-phenyl-3-oxopropyl]phenyl]amino]carbonyl]-,

1,1'-Biphenyl-2-carboxylic acid,2'-[[[4-[3-(3-trifluoromethylphenyl)-1-oxo-propyl]phenyl]amino]carbonyl]-,

1,1'-Biphenyl-2-carboxylic acid,2'-[[[4-[benzyloxy]phenyl]amino]carbonyl]-,

1,1'-Biphenyl-2-carboxylic acid,2'-[[[4-[2-phenylethoxy]phenyl]amino]carbonyl]-,

1,1'-Biphenyl-2-carboxylic acid,2'-[[[4-[4-phenylbutyloxy]phenyl]amino]carbonyl]-,

1,1'-Biphenyl-2-carboxylic acid,2'-[[[4-[2-phenoxy-1-oxoethyl]phenyl]amino]carbonyl]-,

1,1'-Biphenyl-2-carboxylic acid,2'-[[[4-[2-phenyl-2-oxo-ethoxy]phenyl]amino]carbonyl]-.

[1,1'-Biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3,4-dichlorophenyl)propoxy]phenyl]amino]carbonyl]-.

The usefulness of the compounds of the present invention, particularlyas antiasthma and antiallergy agents is demonstrated by theireffectiveness in various standard pharmacological test procedures. Adescription of each procedure follows.

BINDING OF ³ H-LEUKOTRIENE D₄ TO GUINEA PIG LUNG MEMBRANES (RBL)

Materials

[14,15-³ H]leukotriene D₄ (³ H-LTD₄) (25 Ci/mmol and 40 Ci/mmol) ispurchased from New England Nuclear. Unlabeled LTC₄ is a gift of OnoPharmaceuticals (Japan). LTC₄, LTD₄, and LTE₄ are purchased as methylesters from Paesel GmbH (Frankfurt, W. Germany). Concentrations of thePaesel leukotrienes are calculated from their absorbance at 280 nm.Leukotriene esters are saponified overnight under N₂ in 3.3% potassiumcarbonate at room temperature. Tritiated leukotrienes are stored asreceived from New England Nuclear at -20° C. Ono LTC₄ (5 μg/ml) isstored at -60° C. in phosphate buffer pH 6.8. Saponified Paeselleukotrienes are stored at -60° C. in 3.3% potassium carbonate (pH9.0-9.5). Aliquots of leukotrienes are taken from stock solutionsimmediately after thawing, after which the stock solutions areimmediately refrozen. 2-Amino-2-(hydroxymethyl)-1,3-propanediol (Tris)is Sigma pH 7.7 pre-set crystals, and dimethylsulfoxide in Aldrich GoldLabel.

Preparation of crude lung membranes

Two pairs of lungs (1.3 g) from freshly sacrificed 300 g male guineapigs (older animals gave substantially lower binding) from Kuiper RabbitFarm, Gary, IN are disrupted with a Polytron PT 10 (setting 4) for 30seconds in 20 ml ice-cold 50 mM Tris adjusted with HCl to pH 7.7 at 25°C. (Tris buffer), filtered through a single layer of gauze to removeconnective tissue, and centrifuged at 50,000 xg for ten min. The pelletis resuspended by homogenization with a Polytron in 20 ml Tris buffer,centrifuged at 50,000 xg for ten min., resuspended, incubated at 37° C.for 30 minutes, and centrifuged again. The final pellet is resuspendedin Tris buffer and either used fresh or stored at -70° C.

Binding assay

All incubations are in triplicate for 60 minutes at 25° C. in 12×75 mmpolystyrene tubes containing 1 ml Tris buffer with 20 mg original tissuewet weight of guinea pig lung membranes, 0.2 nM ³ H-LTD₄ (6,000-11,000cpm), 10 mM MgCl₂, and 1% dimethylsulfoxide. Leukotrienes are diluted inTris buffer. All other test compounds are dissolved at 10 mM indimethylsulfoxide on the same day as the experiment, and diluted indimethylsulfoxide to 100× the final incubation concentration. Controlincubations receive an equal volume (10 μl) of dimethylsulfoxide; theresulting concentration of dimethylsulfoxide had no effect on binding. ³H-LTD₄ is diluted to 2 nM in Tris buffer. The membrane suspension (20mg/0.89 ml) contains sufficient MgCl₂ to give 10 mM final concentrationin the incubation. For test compounds with IC₅₀ values less than 1 μM,the order of incubations is test compound (10 μl), ³ H-LTD₄ (100 μl),and membranes (0.89 ml). For test compounds with IC₅₀ values greaterthan 1 than 1 μM and limited water solubility, the order of additions istest compound, membranes, and ³ H-LTD₄. All additions are performed at0° C. Immediately after the last addition, the incubation is initiatedby agitating the rack of tubes on a vortex mixer and warming to 25° C.in a water bath. Tubes are vortexed at least once more during theincubation to ensure that the membranes remained suspended. Incubationsare terminated after 60 minutes by filtering under reduced pressurethrough 25 mm Whatman GF/B filters followed by rapid washing three timeswith 4 ml of ice-cold Tris buffer. Filters are added to scintillationvials with 8 ml Formula 947 (New England Nuclear), left overnight,shaken, and the radioactivity counted in a scintillation counter(efficiency 40%). Nonspecific binding, defined as binding of ³ H-LTD₄ inthe presence of 100 nM LTC₄, is 300-500 cpm for all lots of ³ H-LTD₄.Binding to the filters in the absence of tissue is about 100 cpm, and isnot affected by unlabeled LTC₄. Specific binding, defined as totalbinding minus nonspecific binding, varied considerably from lot to lotof ³ H-LTD₄. Specific ³ H-LTD₄ binding ranged from 1500 to 3000 cpm, andis greater than 80% of total binding for the better lots of ³ H-LTD₄.

TO EVALUATE THE EFFECT OF EACH COMPOUNDS AS A 5-LIPOXYGENASE INHIBITORIN COMPARISON TO STANDARD REFERENCE AGENTS IN HUMAN LEUKOCYTES (5LOA1)

The purpose of this assay is to evaluate the activity of each compoundas an inhibitor of human leukocyte 5-lipoxygenase.

Arachidonic acid and calcium ionophore A23187 are obtained from Sigma(St. Louis, MO). Silica gel plates, GF are obtained from Analtech(Newark, DE). Arachidonic acid, (1-¹⁴ C) and 5-HETE (³ H), 5(S)-hydroxy-6-trans, 8,11,14-cis eicosatetraenoic acid, are obtainedfrom New England Nuclear (Boston, MA). Six percent Dextran-70 in 0.9%NaCl is obtained from Cutter Labs (Berkeley, CA).

Preparation of Leukocytes

Fresh blood from normal adult men who had not received any drugs for atleast the previous five days is obtained by the Community ResearchClinic (WL/PD) using venipuncture and collected into heparinizedvacuotainer tubes. To every 100 ml of pooled blood is added 25 ml ofdextran solution (6% dextran -70 in 0.9% sodium chloride containing 3%dextrose) and this is mixed gently in a plastic cylinder. The mixture isleft to stand at room temperature for at least 90 minutes. The upperlayer which is rich in leukocytes and platelets is then carefullydecanted into 50 ml plastic tubes and centrifuged at about 100×g foreight minutes in an IEC centrifuge and rotor number 269 (about 600 rpm).The supernatant fluid is discarded and the pellet is resuspended in 10ml of 0.87% ammonium chloride for exactly two minutes. This procedure isto lyse completely contaminating red blood cells. Leukocytes are thenseparated by centrifugation for ten minutes. The pellet is washed threetimes by suspension in 20 ml PBS (sodium chloride, 7.1 g; Na₂ HPO₄, 1.15g; KH₂ PO₄, 0.2 g, and KCl, 0.2 g/L) and centrifuged as before. Thefinal pellet is suspended in PBS containing 0.87 mM CaCl₂. Viability ofthe cells is then checked using trypan blue exclusion method and isfound to be over 90%.

5-Lipoxygenase Enzyme Assay

Cells in suspension (0.98 ml) are incubated with or without testcompounds for five minutes at 37° C. in a shaking water bath. At thistime a 17 μl mixture is prepared per 1 ml of cell suspension: 100 mMarachidonic acid, 1 μl, 0.05 μCi ¹⁴ C-arachidonic acid in 5 μl; 1 mMcalcium ionophore A23187, 10 μl (1). This mixture is added and theincubation continued for five minutes. The reaction is stopped by addingfour volumes of absolute ethanol and the mixture is kept in ice for 30minutes. The floculated precipitate is separated by centrifugation atabout 37,000×g for 20 minutes (Beckman Instruments rotor number 40). Thealcohol extract is taken to dryness under a stream of nitrogen and theresidue is dissolved in 100-200 μl absolute ethanol. At the time anyturbidity is removed by centrifugation. An aliquot (25-50 μl) is appliedonto 20×20 cm silica gel TLC plate and developed using the followingsolvent system: diethyl ether, petroleum ether (20°-40° C.), acetic acid(50:50:1 v/v). Zones of 1 cm apart are scraped from the TLC plate andtransferred to mini-vials. Methanol (0.5 ml) is added to dissolve theradioactivity adsorbed to the silica gel and scintillation fluid (H. P.,Beckman), 5 ml is then added and vials are counted in a liquidscintillation counter. A sample of ³ H-5-HETE is applied and used forthe identification of the formed 5-HETE.

Total radioactivity in the test as well as the control samples arenormalized and the amount of 5-HETE present is calculated accordingly.

IC₅₀ values are defined as the concentrations of test agents whichcaused a 50% inhibition of the formation of 5-HETE as compared tocontrol and are determined by inspection of the concentration-responsivecurves.

5-LIPOXYGENASE ASSAY USING ISOLATED HUMAN LEUKOCYTES (5LOA2)

The formation of 5-HETE in human leukocytes is considered a measure of5-lipoxygenase activity. The protocol is described in the following.

Fresh heparinized or EDTA treated human blood is mixed with 6%dextran-3% dextrose in isotonic saline in the ratio 0.25 ml dextransolution per 1.0 ml blood. After mixing the blood is allowed to sit atroom temperature for about 90 minutes while the RBC's settle. Duringthis period, the plasma is removed with a plastic pipette to nalgenstubes.

The plasma is centrifuged at 800 rpm (125 kg) on the Beckman Td-brefrigerated centrifuge to remove the platelets (which remain in thesupernatant). The pellet, consisting of Leukocytes and erythrocytes, istreated with 10 ml 0.87% ammonium chloride at room temperature for fourminutes, lysing the red cells. At the end of four minutes the cells arediluted with a 2× volume of phosphate buffered saline, pH 7.4, andcentrifuged for ten minutes. The cells are washed three times with thephosphate buffered saline. Any of the pelleted cell matter which is noteasily resuspended is discarded during the washings--the materialcontains platelets (12-lipoxygenase activity).

After washing, the cells are resuspended in phosphate buffered salinecontaining 1.0 mM calcium and 0.5 mM magnesium. After counting the cellsare diluted to 1.5-2.0×10⁷ leukocytes per milliliter.

To each polypropylene reaction tube is added 0.48 ml leukocytes in Ca-Mgphosphate buffered saline, pH 7.4; 1-5 μl test compound dissolved inDMSO and buffer; or DMSO for control tubes.

The tubes preincubate at 37° C. for five minutes.

The reaction is started by adding 20 μl of the following, 0.5 μl 20 mMarachidonic acid--final concentration=20 μm; 1 μl 5 mM calcium ionophoreA23187--final concentration=10 μm; and 18.5 μl buffer.

The reaction proceeds for five minutes, then is stopped by adding 0.5 ml0.5 mM ice cold Tris buffer, pH 8.0. The tubes are chilled on ice forten minutes and then extracted three times with a total of 3.5 ml ethylacetate (3.0 ml removed).

The tubes can be stored at this point. For extended storage, the tubesshould be filled with nitrogen.

The ethyl acetate is evaporated with a Sorvall Speed-Vac. The residue isdissolved in ethanol. The tubes can also be stored at this point at -20°C. under nitrogen.

A portion of the ethanol solution is injected into the HPLC system for5-HETE quantitation.

The HPLC system consists of Hewlett-Packard 1040A UV spectrophotometrysystem with an HP85 computer. Injections are made automatically with aWaters WISP 710B. The pump is a Spectra Physics SP8700. Peaks aremeasured with a Hewlett Packard 3390A integrator. An RP C-18 column isused. The solvent system is isocratic; the solvent is 70% methanol and30% 0.01M sodium acetate, pH 5.7, pumped at 1.0 ml/min. The flow ismonitored at 235 nm for 5-HETE quantitation. Using a 15 cm AlltechNucleosil C-18 5 μM column provides for a sample turnaround time ofabout 16 minutes.

IC₅₀ is calculated as the amount of test agent that causes 50%inhibition of the formation of 5-HETE relative to the control.

SMOOTH MUSCLE RECEPTOR AGONISM--ANTAGONISM ACTIVITY EVALUATIONS IN VITRO(IVAS) ISOLATED GUINEA PIG LUNG PARENCHYMA

Isolated organs from experimental animals, have long been used tocharacterize the mechanism of drug activity and to elucidate theintrinsic potency and specificity of this action. The use of isolatedtissues, kept viable in an oxygenated and nutritive tissue bath, hasmost often been used to measure specific drug activity on musclecontractile processes including the associated electrical phenomenon.The agonistic and antagonistic activity of a test drug on specializedtissue areas (receptors) associated with muscle contraction, especiallyin cardiovascular and visceral smooth muscle, have, in recent years,received considerable attention by many investigators.

It is generally agreed that the final decisive proof of therapeuticpotential resides in the level of activity and safety of the drug in theintact animal. However, such information concerning drug action isderived prior to these final tests in animals by the characterization ofthis action in the isolated system which is unhindered by variability indrug absorption, distribution, and metabolism. The potency of a testcompound for receptor stimulation or antagonism, for example, can easilybe determined since the concentration of the compound to which themuscle cell, or specialized responsive area on this cell, is exposed canbe easily controlled and correlated with the recorded activity of musclecontraction or relaxation.

The procedures described herein aare specifically concerned with thedefinition of a test drug action on isolated lung strips. Drug action ischaracterized either by an action per se on the tissues or byinteraction with known smooth muscle receptor stimulants or antagonists.

Usually, the tests will be used to evaluate the ability of a testcompound to antagonize the contraction induced by standard agonists inthis case LTC₄. A relative potency is derived by comparison with one ofthe standard blocking drugs. In caution, relative potencies of testcompounds to known standards are valid only if full dose response curvesare generated and the compound compared generate parallel dose/responsecurves with common maxima. Appropriate "within-trial" comparisons arealso necessary.

Molecular antagonists compete with the agonist for the receptor. Anexample of molecular antagonism would be the antihistamine activity ofmepyramine relaxing a contraction of tissue produced by histamine. Aphysiological antagonist produces a response by action on a differentreceptor that causes an effect in opposition to the agonism. An exampleof physiological antagonism would be relaxation by the adrenoreceptoragonist isoproterenol of a tissue contraction procedure by histamine orcarbachol.

Materials and Equipment

A permanent isolated bath system is used which facilitates the verticalsuspension of a segment of tissue in small volume of a specificnutritive buffer. Frittered glass terminals provide dispersed bubbles of95% oxygen and 5% carbon dioxide at the base of the individual tissuebaths and the buffer reservoir. All baths and the reservoir aredouble-jacketed and water at 37° C. is circulated through the jacket ofthe baths to provide stable temperature regulation. The baths arearranged so that fresh solution from the reservoir passes through ajacketed glass coil to the base of the bath. The bubbling gas stirs thebath solution. During equilibration, fresh solution enters and excesssolution is allowed to overflow. Unless a particular drug is requiredthroughout the experiment and it is incorporated in the buffer,individual drugs are introduced directly into the bath by longhypodermic needles to the base of the bath without disturbing thetissue. Drugs are removed by rapidly draining the tissue bath from thebottom and refilling. Tissues are washed a minimum of six times. Theemptying and refilling sequence takes only five seconds.

The contraction and relaxation of the smooth muscle preparation can berecorded either isotonically (change in length) or isometrically (changein tension). Isometric transducers are more sensitive and proportional.Isometric contractions are recorded electronically using Grass FT 0.03Cforce displacement transducers and Beckman Dynagraph recorders. Thetissue is suspended vertically with one end of the segment anchored tothe bottom of the bath and the other end attached to the mechanicalelectrical transducer muscle level. The electrical output of thistransducer moves a pen on a moving chart in proportion to the forcedisplacement.

Isolated Strip of Lung

Lung tissue is obtained from normal male guinea pigs. The heart andlungs are removed as a unit, placed in buffer, and the lungs areperfused with buffer by the spontaneously beating heart for severalminutes. Distal strips of lung from the diaphragmatic lobe,approximately 0.3 cm wide and 3 cm long are removed and attached toGrass strain gage transducers. The desirable preload on the lung stripsis 0.3 g of tension, with the sensitivity at 0.02 mv/cm on the amplifierand 0.5 mv/cm on the preamp, and the calibration of the chart is 1 mm=5mg force displacement. After equilibration and priming, the tissue isready for drug assay. When contracted with histamine, the contractionreaches the maximum within two minutes with a steady decline which takesover an hour to approach baseline. A threshold dose of histamine isapproximately 3×10⁻⁶ M. At 3×10⁻⁵ M, histamine produces a contractionapproximately 75% of maximum. For the leukotriene C₄, the contractionhas a short slower ascension with a decline to a plateau afterapproximately seven minutes.

Test Procedures

The potency of the test compound as an antagonist is determined for adefined concentration by adding LTC₄ at hourly intervals for threecontractions in the absence of test compound. Then drug is added and tenminutes later LTC₄ is again added. The inhibition of the contraction isdetermined by comparing the before and after drug responses.

Tissues are primed twice at a lower concentration of agonist (1 ng/mlLTC₄) (a₁) that gives a response (r₁) approximately 15-20% of maximum.The agonist is then increased to a concentration (2 ng/ml LTC₄) (a₂)which gives a response (r₂) approximately 30% of maximum. After responser₂ is obtained in triplicate and the tissue is washed until the tensionreturns to baseline, test compound at a defined concentration is addedto the tissue bath. After monitoring the effect of the drug on baselinetension for ten minutes, agonist at concentration a₂ is given. Theresponse (R) to agonist at concentration a₂ in the presence of testcompound is compared to the previous response (r₂) without testcompound.

INDUCTION AND QUANTITATION OF THE SLOW-ONSET (NONHISTAMINE) BRONCHOSPASMINDUCED BY ANTIGEN CHALLENGE IN SENSITIZED GUINEA PIGS (SRSPIG)

This guinea pig in vivo model was developed to quantitate the pulmonaryeffects of nonhistamine mediated, antigen-induced anaphylaxis.Mepyramine pretreatment is used to block the effects of histamine topermit quantitation of the effects of other biologically activemediators. The method is designed to detect drugs which inhibit therelease or synthesis "other" mediators, such as leukotriene,thromboxane, etc., or drugs blocking the end organ effects of thesemediators. When a sensitized guinea pig is exposed to the sensitizingantigen, it develops a systemic anaphylactic response characterized byincreased pulmonary resistance and decreased dynamic compliance (CDYN).Pulmonary resistance is a measure of the changes in transpulmonarypressure and respiratory flow. Dynamic compliance is an expression ofthe relative changes in tidal volume and transpulmonary pressure. Inthis model, dynamic compliance is used to measure changes in the elasticpropertie of the small airways and pulmonary resistance to measureconstrictive responses of the large airways. In human asthma, smallairway changes appear to be more closely related to the pathogenesis ofdisease. Thus, dynamic compliance effects will be weighted more heavilyin our analysis of the changes produced by drugs. In the presence ofmepyramine, the effects of histamine are blocked. The observed pulmonaryeffects are thought to be largely leukotriene mediated. In the guineapig, thromboxanes may also be involved (1,2) and might augment theleukotriene response. Since leukotrienes are not preformed mediators andtheir effects are slow in onset, in the absence of the histamine effect,a bronchospasm is not seen until approximately 1.5-3 minutes afterantigen administration. However, the leukotriene-mediated response islong lasting.

Materials and Methods

Animals: Male Hartley strain guinea pigs, weighing between 200 and 250 gon delivery, were used. The animals were supplied by the Charles RiverCo., Wilmington, MA.

Sensitization Procedure: Ovalbumin (2×recrystallized, Miles LaboratoriesLtd.) is dissolved (0.2 mg/ml) in saline. The guinea pigs are given asingle intraperitoneal injection of 0.5 ml of the stock solution tosensitize them. The animals are maintained normally for four to sixweeks before use.

Anesthesia: Surgical anesthesia is induced by giving an IP injection ofvalium, 5 mg/kg, followed immediately by Innovar-Vet (Pitman-More), 0.7ml/kg (IM). Each ml of Innovar-Vet contains 0.4 mg fentanyl, 20 mgdroperidol, 1.8 mg methylparaben, and 0.2 mg propylparaben.

Surgical Preparation: The anesthetized guinea pig is shaved in theventral neck region and on the right side of the thorax and affixedsupine to a dissecting board. A midline incision is made in the neck andthe trachea is exposed. The jugular vein is cannulated with PE 50 tubingto administer drugs and antigen. The trachea is cannulated with PE 240tubing. If the protocol required measuring blood pressure, the carotidartery is isolated and cannulated with PE 50 tubing. A spear-likecannula is then inserted into the thorax to measure transpulmonarypressure. Needle electrodes are inserted into the musculature of eachleg to record electrocardiograms.

Pulmonary Mechanics Methods: The tracheal cannula is connected inline toa Fleisch 000 pneumotachograph (factory calibration 10 cm H₂ O=22.2ml/sec) and a Validyne differential pressure transducer, to measurerespiratory flow. Transpulmonary pressure is measured by connecting oneside of a Validyne pressure transducer to the side arm of the trachealcannula and the other side to the cannula inserted into the thorax atapproximately the 5th intercostal space.

The pressure and flow signals are fed into Validyne preamplifiers andthen transmitted to a pulmonary mechanics computer (Buxco). The computercomputer calculates total pulmonary resistance, dynamic compliance,tidal volume, respiratory rate, and minute volume. An arterial pressuretransducer (Statham P23) connected directly to the pulmonary computer isused when monitoring arterial blood pressure. The computer calculatessystolic blood pressure, diastolic blood pressure, mean blood pressureand heart rate from the arterial blood pressure signal. The analogsignals of the respiratory and cardiovascular parameters are also fedinto a data logger (Buxco Model DL-12) which digitized the signals foroutput to a electric typewriter (TI #700). Simultaneously, the analogsignals are fed into an eight 8 channel recorder (Beckman Dynograph) tomake permanent experimental records.

Animals are mechanically ventilated using a rodent respirator (Harvard#680). In order to monitor pump pressure, a pressure transducer(Statham, Model PM131TC) is connected by a T-tube to the output side ofthe pump. The signal is sent directly to the Beckman recorder.

Experimental Protocol: The animal is anesthetized as described above,surgically prepared and connected to the appropriate monitoring devices.In animals that are dosed intraperitoneally, the drug is administered 30minutes prior to antigen challenge. The animal is then attached to therespirator and the stroke volume adjusted to deliver 2.5 ml/breath at arespiratory rate of 65 breaths/min. The animal is given 1.2 mg/kg (IV)of succinyl choline (1) to arrest spontaneous breathing. One minutelater, mepyramine, 2.0 mg/kg, is administered intravenously. Six minutesafter the mepyramine, ovalbumin is infused over a one-minute period andthe animal monitored for 15 minutes. The dose of ovalbumin is selected(3-10 mg/kg) to produce a decrease in CDYN five minutes after ovalbuminto approximately 35%±15 of the baseline value. six minutes later, allanimals are dosed with the beta adrenoreceptor antagonist propranolol,2.5 mg/kg.

Calculations and Rating System: The "baseline" values are determined byaveraging the three one-minute averages of CDYN values immediately priorto ovalbumin administration. The five-minute postovalbumin % of baselineCDYN (mean ±SE) for each animal and the means for both control (A) anddrug-treated groups (B) are calculated. Percent inhibition is thencalculated using the formula: ##EQU1## Results

After anitigen challenge, untreated animals develop a bronchospasm.There is usually a delay of 1.5-3 minutes in the onset of the responseto antigen challenge, after which, there is a rapid decline in dynamiccompliance (usually to 20-50% of baseline) and an increase in pumppressure, transpulmonary pressure, and pulmonary resistance. Tidalvolume might decrease slightly, but because the animal is respiratedthis decrease is usually small. The bronchospasm reaches its peak atfour to five minutes and is stable for more than 15 minutes if no othertreatments are given. The propranolol is given after six minutes toblock any beta sympathomimetic tone and/or the effects of catecholrelease. This elicits a maximal response. Thus, animals with highsympathetic activity develop a more intense bronchoconstrictionfollowing propranolol.

HISTAMINE RELEASE FROM HUMAN BASOPHILS (HHB) PROCEDURE OF QUANTITATINGACTIVE HISTAMINE RELEASE, AND ITS INHIBITION BY DRUGS, FROM BASOPHILS OFHUMAN BLOOD USING AN AUTOMATED FLUOROMETRIC HISTAMINE ASSAY

Mediator (histamine) release from human blood basophils is an assaywhich provides a model for evaluating potential antiallergy (mediatorrelease inhibitors) compounds.

Blood from allergic donors is obtained from volunteers through theCommunity Research Clinic. The leukocytes are removed after Hespan®sedimentation, washed and concentrated using calcium and magnesium freeHepes buffered saline and resuspended in Hepes Hepes buffered salinewith 1 mM calcium and magnesium added back. The cells are incubatedbriefly at room temperature with either buffer alone (for measuringspontaneous release and challenge agent stimulated release) 6%perchloric acid (for measuring total available histamine in the cells)or drug at various concentrations. The cells are then either treatedwith buffer alone (to measure control spontaneous and drug effect onspontaneous release) or appropriate challenge agent (e.g., anti-IgE,antigen) mixed and incubated for 45 minutes at 37° C. in a shaking waterbath. The reaction is stopped by centrifugation and the supernate ispoured into autoanalyzer sample cups and samples assayed for assayed forhistamine content using an automated fluorometric assay. By comparingthe histamine release of drug-treated cells and the challenge agentcontrols, drug inhibition of mediator (histamine) release can beevaluated.

Histamine release is induced by (1) an aqueous antigen extract (shortragweed or house dust), or (2) anti-IgE antisera. Whole human blood isobtained from volunteers through the Community Research Clinic (CRC-92).Volunteers are chosen on the basis of adequate histamine release inducedby antigen, or antiIgE antisera challenge. Leukocytes are incubated invitro with the challenge agents at several concentrations. Dose-responsecurves for each volunteer, using each challenge agent, are generated.Test compounds are evaluated for inhibition of a near maximal responseto one or more challenge agents. By including appropriate controls, itis possible to measure the effect of the test compound, at variousconcentrations, in several ways: (1) the effect of the compound onhistamine release for each challenge agent; (2) the effect of the testcompound on spontaneous release of histamine in the absence of thechallenge agent (buffer alone); and (3) the effect of the test compoundon the histamine assay.

Histamine is assayed using an automated continuous flow system whichquantitates samples of 0.4 ml or more in volume, and concentrations ofapproximately 0.5 ng/ml or more. The rate of analysis is 30samples/hour.

Methods of preparing leukocytes for this assay are described by Theuson,et al, J. of Immunology, Vol. 123, p. 426 (1979).

Protocol Design

Samples are run in triplicate, using either 1.5 ml polypropylene cappedreaction tubes, or 5.0 ml plastic uncapped tubes. Test compounds andchallenge agents are prepared in HACM buffer, as described above. Fixedvolume pipettes are used.

Test compound or vehicle control is added to three reaction tubes at1.5× the final desired concentration (i.e., 400 μl of test compound per600 μl total reaction volume). One hundred μl of cells is added to eachtube and the mixture is incubated for eight minutes at room temperature,and two minutes at 37° C. before antigen or other stimulus challenge.One hundred μl of the challenge agent at 6× the final concentration isthen added, and the final mixture is incubated at 37° C. for 45 minutesin a shaking water bath. This ensures that the cell preparation isconstantly in suspension. The reaction is stopped by centrifugation at2000 RPM for three minutes at 4° C. The supernate (≅500 μl) is pouredinto 2.0 ml autoanalyzer beakers and assayed for histamine by thefluorometric method.

In each experiment, cells from one donor are challenged with one or moreof the challenge agents, according to the designed protocol and thepreviously determined sensitivity of the donor to particular challengeagents. Short ragweed and house dust concentrations are expressed inPNU/ml, and anti-IgE antisera in dilutions, e.g., 1E-5 (1:100,000), 3E-5(1:30,000), and 1E-4 (1:10,000).

Calculation and Interpretation of Results

The total histamine concentration in the "total" (acid-treated) samplesmust be 15 ng/ml to be acceptable. Spontaneous release of histamine fromthe cells should not exceed 15% of the total histamine, and isfrequently <5%. The maximum percentage histamine released varies withthe donor. The net amount released by the challenge agent must exceed25% of the total cellular histamine to confidently assess inhibition bytest compounds. Spontaneous histamine release is subtracted from both"totals" and challenged cells to calculate net percent release. Percentinhibition is calculated using the following formula: ##EQU2## TheAutomated Analyzer

The fluorometric method for histamine determination was originallydescribed by Shore, et al² and has been modified to increase both itsspecificity and sensitivity. See R. P. Siraganian, J. of Immunol. Res.,Vol. 17, p. 283 (1975).

When tested by the above described procedures and shown by the notationsunder the acronym of each test, various compounds of the Formula I asdefined above indicated activity as shown in Table 1.

                                      TABLE 1                                     __________________________________________________________________________                   33 μM cpd                                                                          33 μM cpd.                                                                          IC.sub.50                                                                            IC.sub.50                              Example Number                                                                         IC.sub.50 RBL1                                                                      HHB (antigen)                                                                         HHB (Tripeptide)                                                                       5LOA (100)*                                                                          5LOA (20)*                                                                           SRS-PIG                                                                            IVAS                       __________________________________________________________________________     1       1.0   72      72       9.6    6.7    51%  90% @ 3 μM               2       4.6   12      96                     64%                              3       4.8   94      93       4.6           16%  IC.sub.50 = 1.0             4       8.5   101     97       9.3            3%  A @ 10                      5       14.5  72      83       40.0                                           6       1.0                                       A @ 10                      7       88.7  -5      93       >20    >20                                     8       28.7  67      111      >20                                            9       6.5   87      100             19                                     10       9.7   62      74                                                     11       13.5                    12                                           12       13.7  -42     99                                                     13       5.4                                                                  14       3.7   41      96                                                     15       4.2   @ 10 is 39                                                                            -17             4.6                                    16       4.3           84              22     33%                             17       10.5          97                                                     18       16.5  60      98                                                     19       6.7   59      90              7.2                                    20       11.2                                                                 21       2.3   91      93                     21%                             22       10.6   30*    100                                                                   ragweed                                                        23       50.2  -8      45                                                     24       10.5  45      98                                                     25       14.5  26      89                                                     26       36.0  -10     65                                                     27        0.23 lysis   lysis                                                  28       5.6   85      86                                                     29       5.0   lysis   lysis                                                  30       1.2   101     94                                                     31       2.2                                                                  32       14.7                                                                 33       2.4                                                                  34       6.5                                                                  35       30.7                                                                 36       100.0                                                                37       --                                                                   38       >100                                                                 39       --                                                                   40       >100                                                                 41       56.0                                                                 42       >100                                                                 43       33.6                                                                 __________________________________________________________________________     *Concentration of exogenous arachidonic acid (μM)                     

Accordingly, the present invention also includes a pharmaceuticalcomposition for treating asthma or allergies comprising an effectiveamount of a compound of the Formula I as defined above together with apharmaceutically acceptable carrier.

The present invention further includes a method for treating asthma orallergies in mammals suffering therefrom comprising administering tosuch mammals either orally or parenterally a correspondingpharmaceutical composition containing a compound of Formula I as definedabove in appropriate unit dosage form.

For preparing pharmaceutical compositions from the compounds describedby this invention, inert, pharmaceutically acceptable carriers can beeither solid or liquid. Solid form preparations include powders,tablets, dispersible granules, capsules, cachets, and suppositories. Asolid carrier can be one or more substances which may also act asdiluents, flavoring agents, solublizers, lubricants, suspending agents,binders or tablet disintegrating agents; it can also be encapsulatingmaterial. In powders, the carrier is a finely divided solid which is inadmixture with the finely divided active compound. In the tablet theactive compound is mixed with carrier having the necessary bindingproperties in suitable proportions and compacted in the shape and sizedesired. The powders and tablets preferably contain from 5 or 10 toabout 70 percent of the active ingredient. Suitable solid carriers aremagnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin,dextrin, starch, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose, a low melting wax, cocoa butter, and the like.The term "preparation" is intended to include the formulation of theactive compound with encapsulating material as carrier providing acapsule in which the active component (with or without other carriers)is surrounded by carrier, which is thus in association with it.Similarly, cachets are included. Tablets, powders, cachets, and capsulescan be used as solid dosage forms suitable for oral administration.

For preparing suppositories, a low melting wax such as a mixture offatty acid glycerides or cocoa butter is first melted, and the activeingredient is dispersed homogeneously therein as by stirring. The moltenhomogeneous mixture is then poured into convenient sized molds, allowedto cool and thereby to solidify.

Liquid form preparations include solutions, suspensions, and emulsions.As an example may be mentioned water or water propylene glycol solutionsfor parenteral injection. Liquid preparations can also be formulated insolution in aqueous polyethylene glycol solution. Aqueous solutionssuitable for oral use can be prepared by dissolving the active componentin water and adding suitable colorants, flavors, stabilizing andthickening agents as desired. Aqueous suspensions suitable for oral usecan be made by dispersing the finely divided active component in waterwith viscous material, i.e., natural or synthetic gums, resins,methylcellulose, sodium carboxymethylcellulose, and other well-knownsuspending agents.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions, and emulsions. These particular solid form preparations aremost conveniently provided in unit dose form and as such are used toprovide a single liquid dosage unit. Alternately, sufficient solid maybe provided so that after conversion to liquid form, multiple individualliquid doses may be obtained by measuring predetermined volumes of theliquid form preparation as with a syringe, teaspoon, or other volumetriccontainer. When multiple liquid doses are so prepared, it is preferredto maintain the unused portion of said liquid doses at low temperature(i.e., under refrigeration) in order to retard possible decomposition.The solid form preparations intended to be converted to liquid form maycontain, in addition to the active material, flavorants, colorants,stabilizers, buffers, artificial and natural sweeteners, dispersants,thickeners, solubilizing agents, and the like. The liquid utilized forpreparing the liquid form preparation may be water, isotonic water,ethanol, glycerine, propylene glycol, and the like as well as mixturesthereof. Naturally, the liquid utilized will be chosen with regard tothe route of administration, for example, liquid preparations containinglarge amounts of ethanol are not suitable for parenteral use.

Preferably, the pharmaceutical preparation is in unit dosage form. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, for example, packeted tablets, capsules, and powders invials or ampoules. The unit dosage form can also be a capsule, cachet,or tablet itself or it can be the appropriate number of any of these inpackaged form.

The quantity of active compound in a unit dose of preparation may bevaried or adjusted from 0.1 mg to 500 mg preferably to 1 to 10 mgaccording to the particular application and the potency of the activeingredient. The compositions can, if desired, also contain othercompatible thereapeutic agents.

In therapeutic use as described above, the dosages may be varieddepending upon the requirements of the patient, the severity of thecondition being treated, and the compound being employed. Determinationof the proper dosage for a particular situation is within the skill ofthe art. Generally, treatment is initiated with smaller dosages whichare less than the optimum dose of the compound. Thereafter the dosage isincreased by small increments until the optimum effect under thecircumstances is reached. For convenience, the total daily dosage may bedivided and administered in portions during the day if desired. ##STR1##

We claim:
 1. A compound having the formulawherein R₁ is hydrogen orlower alkyl of from one to four carbons, inclusive; B is (a) --(CH₂)_(m)-- (b) --C(O)--CH═CH-- (c) --C(O)--(CH₂)_(q) -- (d) --(CH₂)_(q) --C(O)--(e) --O--(CH₂)_(q) -- (f) --O--(CH₂)_(m) --O-- (g) --CH(OH)(CH₂)_(q) --(h) --(CH₂)_(q) CH(OH)-- (i) --CH═CH--C(O)-- (j) --(CH═CH)_(m) -- (k)--(CH₂)_(q) --O--wherein m is an integer from one to six and q is aninteger from zero to six and Z is ##STR2## wherein n is and integer ofzero to five and, R may be the same or different when n is two or moreand is lower alkyl of from one to four carbons, hydroxy, alkoxy of fromone to four carbons, inclusive, inclusive, halogen, or Z as definedabove with the proviso that R may only be attached to a carbon and thateach carbon atom may carry only one R, or pharmaceutically acceptablesalts thereof; R₁ is lower alkyl up to butyl.
 2. A compound of claim 1wherein B is --(CH₂)_(m) -- and is attached to the position three orfour of the phenylamido group in the compound of Formula I as definedabove.
 3. A compound of claim 1 wherein B is --C(O)--(CH₂)_(q) -- and isattached to the three or four position of the phenylamido group of thecompound of Formula I as defined above.
 4. A compound of claim 1 whereinB is --O--(CH₂)_(q) --.
 5. A compound of claim 1 wherein B is--O--(CH₂)_(m) --O--.
 6. A compound of claim 2 wherein m is one throughthree and Z is ##STR3## wherein n is zero or n is two and R is halo oralkyl of from one to four carbons, inclusive.
 7. A compound of claim 3wherein q is one through three and Z is ##STR4## wherein n is zero or nis two and R is halo or alkyl of from one to four carbons, inclusive. 8.A compound of claim 4 wherein q is one through three and Z is ##STR5##wherein n is zero or n is two and R is halo or alkyl of from one to fourcarbons, inclusive.
 9. A compound of claim 5 wherein m is one throughthree and Z is ##STR6## wherein n is zero or n is two and R is halo oralkyl of from one to four carbons, inclusive.
 10. A compound of claim 7wherein m is three and n is zero or n is two and R is chloro or methyl.11. A compound of claim 7 wherein q is two or three and n is zero or nis two and R is chloro or methyl.
 12. A compound of claim 8 wherein q isthree and n is zero or n is two and R is chloro or methyl.
 13. Acompound of claim 9 wherein m is two or three and n is zero or n is twoand R is chloro or methyl.
 14. A compound of claim 10 wherein theembodiment is [1,1'-biphenyl]-2-carboxylic acid,2-[[[4-[3-(3,4-dichlorophenyl)propyl]phenyl]amino]carbonyl]-.
 15. Acompound of claim 10 wherein the embodiment is[1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-(3-phenyl)]propyl]phenyl]amino]carbonyl]-.
 16. A compound ofclaim 11 wherein the embodiment is [1,1'-biphenyl]-2-carboxylic acid;2'-[[[4-[3-(3,4-dichlorophenyl)-1-oxopropyl]phenyl]amino]carbonyl]-. 17.A compound of claim 11 wherein the embodiment is[1,1'-biphenyl]-2-carboxylic acid;2'-[[[3-[3-(3,4-dichlorophenyl)-1-oxopropyl]phenyl]amino]carbonyl]-. 18.A compound of claim 11 wherein the embodiment is[1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-(1-oxo-3-phenylpropyl)phenyl]amino]carbonyl]-.
 19. A compound ofclaim 11 wherein the embodiment is [1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-[3-[3-(1,3-benzodioxol-5-yl)-1-oxopropyl]phenyl]amino]carbonyl]-.20. A compound of claim 12 wherein the embodiment is[1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3-phenylpropoxy)]phenyl]amino]carbonyl]-.
 21. A compound ofclaim 12 wherein the embodiment is [1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-[3-(3,4-dichlorophenyl)propoxy]phenyl]amino]carbonyl]-.
 22. Acompound of claim 13 wherein the embodiment is[1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-[3-[3,4-dichlorophenoxy]propoxy]phenyl]amino]carbonyl]-.
 23. Acompound of claim 13 wherein the embodiment is[1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-[2-(2,5-dimethylphenoxy]ethoxy]phenyl]amino]carbonyl]-.
 24. Acompound of claim 13 wherein the embodiment is[1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-[2-(3,4-dichlorophenoxy)ethoxy]phenyl]amino]carbonyl]-.
 25. Acompound of claim 13 wherein the embodiment is[1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-(2-phenoxyethoxy)phenyl]amino]carbonyl]-.
 26. A compound ofclaim 13 wherein the embodiment is [1,1'-biphenyl]-2-carboxylic acid,2'-[[[4-[3-phenoxypropoxy)phenyl]amino]carbonyl]-.
 27. A compositionhaving compounds of claim 1 and a pharmaceutically acceptable carrier inunit dosage form.
 28. A method of treating asthma or allergy byadministering to a mammal suffering therefrom an antiasthma orantiallergy effective amount of a compound of claim
 1. 29. A method oftreating an immunoinflammatory condition by administering to a mammalsuffering from the condition an antiimmunoinflammatory effective amountof a compound of claim
 1. 30. A method of treating cardiovasculardisease by administering to a mammal suffering therefrom acardiovascular effective amount of a compound of claim
 1. 31. A methodof treating migraine by administering to a human suffering therefrom anantimigraine effective amount of a compound of claim 1.